N- and O-Glycosylation Site Analysis Services
N- and O-glycosylation site analysis is designed to identify specific glycosylation sites within proteins, clarify their modification types and positional distribution, and serves as a key approach to understanding glycoprotein structure and function. N-glycosylation primarily occurs on asparagine (Asn) residues, typically within the conserved Asn-X-Ser/Thr motif. In contrast, O-glycosylation commonly modifies serine (Ser) or threonine (Thr) residues, without a consistent sequence pattern. These modifications influence protein folding, stability, cellular localization, and biological activity, making them essential targets in biopharmaceutical development, biomarker discovery, and glycoscience research.
MtoZ Biolabs offers N- and O-Glycosylation Site Analysis Services through high-resolution mass spectrometry combined with multi-strategy sample preparation workflows, allowing researchers to accurately identify glycosylation sites and explore the structural and functional significance of glycoproteins.
Analysis Workflow
Our N- and O-Glycosylation Site Analysis Services utilize a site-level analysis workflow that combines experimental precision with advanced data interpretation to ensure accurate and reliable identification of glycosylation sites.
1. Protein Digestion
Target proteins are digested using specific enzymes (such as trypsin) to generate peptides containing potential glycosylation sites.
2. Glycopeptide Enrichment and Control Setup
Glycopeptides are enriched, and untreated samples are retained as non-glycosylated peptide controls.
3. LC-MS/MS Analysis
Parallel analysis of peptides and glycopeptides is conducted using high-resolution mass spectrometry platforms such as Orbitrap or Q Exactive.
4. Data Comparison and Occupancy Calculation
Using database search and diagnostic ions, glycopeptide and non-glycopeptide peak areas are compared to calculate the modification occupancy at each site.
5. Data Collation and Bioinformatics Analysis
Visualized charts, modification site distribution maps, and differential analysis results are provided.
Dang, L. et al. Trends Analyt Chem. 2019.
Figure 1. Workflow for N-Glycosylation Site Identification
Why Choose MtoZ Biolabs?
✅ High-Performance Mass Spectrometry Platforms: Equipped with Orbitrap Fusion Lumos, Q Exactive HF, and integrated Nano-LC systems, we deliver high-resolution, high-sensitivity detection of both N- and O-linked glycopeptides.
✅ Versatile Enrichment Strategies: We apply multiple enrichment techniques including lectin affinity, HILIC, and ZIC-HILIC to selectively isolate N- and O-glycopeptides, enhancing detection efficiency even in complex matrices.
✅ Broad Sample Compatibility: Our workflows are optimized for a wide range of sample types, including recombinant proteins, therapeutic antibodies, serum, plasma, cell lysates, and tissue extracts.
✅ Customized Project Solutions: We provide flexible and tailored service packages designed around specific research objectives such as glycan antigen discovery for vaccine development or mechanistic studies of glycosylation regulation. Expert support is available at every stage of the project.
✅ One-Time-Charge: Our pricing is transparent, no hidden fees or additional costs.
Sample Submission Suggestions
To ensure high-quality results in glycosylation site analysis, clients are advised to submit samples with high purity and minimal contamination. We accept various biological sample types including purified proteins, cell lysates, tissue extracts, and serum or plasma.
Samples should be free from salts, detergents, and preservatives. Repeated freeze-thaw cycles and exposure to high temperatures should be avoided to maintain sample integrity. Solid samples such as tissues or cell pellets should be stored frozen, while liquid samples should be shipped with cold packs under low-temperature conditions.
Detailed sample preparation and shipping instructions will be provided by MtoZ Biolabs upon project consultation. We encourage clients to discuss experimental design, sample origin, and target goals with our technical team prior to sample submission to ensure successful downstream analysis.
FAQ
Q1: How to distinguish N-glycosylation and O-glycosylation sites?
N-glycosylation usually occurs within the conserved Asn-X-Ser/Thr motif and can be identified using mass spectrometry combined with database searching. O-glycosylation lacks fixed sequence patterns and requires derivatization or selective enzymatic treatment to improve localization specificity. We employ multi-dimensional approaches to ensure accurate and comprehensive site identification.
Q2: Is glycan structure information included in the analysis?
Upon request, we can annotate glycan structures at identified sites using multistage mass spectrometry and exoglycosidase treatment to reveal glycan composition, linkage patterns, and modification types such as sialylation and fucosylation.
What Could be Included in the Report?
1. Comprehensive Experimental Details
2. Materials, Instruments, and Methods
3. Site-specific Glycosylation Identification Report
4. Data Analysis, Preprocessing, and Estimation
5. Optional Bioinformatics Insights
6. Raw Data Files
MtoZ Biolabs’ N- and O-Glycosylation Site Analysis Services are widely applicable to glycoprotein structural and functional studies, biopharmaceutical characterization, vaccine glycan antigen development, and congenital glycosylation disorder research. We provide high-resolution and high-confidence data on glycosylation site localization and structure, supporting multi-dimensional exploration of glycan biology and potential applications.
For customized service plans or quotation requests, please contact our expert team at any time.
Related Services
N- and O-Glycosylation Site Occupation Analysis Services
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